CN1988926B - Adsorber for direct blood perfusion filled with adsorbent from which water-insoluble fine particles were removed, and method of obtaining adsorbent for direct blood perfusion from which water-insoluble fine particles were removed - Google Patents

Abstract

A direct hemoperfusion adsorber that comprises a water insoluble support having a relatively easily procurable particle diameter distribution without the need to install special granulation equipmentand coating equipment, and that is packed with an adsorbent having water insoluble microparticles removed therefrom; and a method of obtaining a direct hemoperfusion adsorbent having water insoluble microparticles removed therefrom. A direct hemoperfusion adsorber packed with an adsorbent of 300 to 600 m number-basis average particle diameter and 10 to 20% variation coefficient of particle diameter distribution having water insoluble microparticles removed to a safe level therefrom is obtained without any adsorbent loss by the use of a water insoluble microparticle removing mesh of 1.3 to 1.5opening size ratio to the mesh opening size of adsorber.

Description

Adsorber for direct blood perfusion filled with adsorbent from which water-insoluble fine particles were removed, and method of obtaining adsorbent for direct blood perfusion from which water-insoluble fine particles were removed

technical field

The present invention relates to an adsorber for direct blood perfusion filled with an adsorbent having an average particle diameter of 300 μm to 600 μm calculated on the basis of the number of particles, a variation coefficient of particle diameter distribution of 10% to 20%, and water-insoluble fine particles removed . A method for obtaining an adsorbent for direct blood perfusion from which water-insoluble particles have been removed.

Background technique

At present, blood purification adsorption therapy, which actively removes the causative substances present in the patient's blood, has been vigorously tried, and it has been found that it is effective for many difficult diseases. Blood purification adsorption therapy includes plasma perfusion and direct blood perfusion. The plasma perfusion method is a two-stage system of a plasma separation device and a blood purification device, which requires a dedicated device for safe operation, and requires high treatment costs and complicated operations. On the other hand, the direct blood perfusion method is a one-stage system that directly perfuses blood into the adsorber, so the operation is simple, and the amount of blood in extracorporeal circulation is small, which can reduce the burden on the patient. However, one of the problems of the adsorber used in the direct blood perfusion method involves the water-insoluble fine particles present in the adsorber.

The so-called water-insoluble particles are mainly particles from the adsorption material, which can theoretically flow out of the adsorber and enter the body during direct blood perfusion treatment. When water-insoluble fine particles flow into the body in large quantities, they may clog blood vessels or accumulate in organs, which may pose a major safety problem.

Among the adsorbers for direct blood perfusion, the following adsorbers are representative examples of conventional commercially available products (see Non-Patent Document 1).

One of them is an adsorber for removing β2 microglobulin (molecular weight: about 12,000) produced by Chung Yeon Chemical Industry Co., Ltd. The adsorbent used in this adsorber (see Non-Patent Document 2) has a particle diameter of about 500 μm and a uniform particle diameter distribution. Uses It is suitable for amyloidosis accompanied by arthralgia and used during hemodialysis. Since the particle size distribution of the adsorbent is uniform, water-insoluble fine particles can be easily removed without loss of the adsorbent when using a water-insoluble fine particle removing device (see Patent Document 1) or the like. That is, as a countermeasure against water-insoluble fine particles, an adsorbent having a uniform particle diameter distribution is used. However, the production of an adsorbent with uniform particle size distribution (see Patent Document 2) requires a special granulation device (see Patent Document 3). However, the actual situation is that a general-purpose water-insoluble carrier having a uniform particle size distribution suitable as an adsorption material carrier does not exist as a commercial product.

The other is Kuraray Medical Co., Ltd.'s adsorption-type blood purifier for renal assistance. The particle size of the adsorbent used in this adsorber (see Non-Patent Document 2) is 400-900 μm, and it is used as a kidney aid for removing uretoxin-causing substances (molecular weight: about 100-2000). In order to be used as an adsorbent for direct blood perfusion, polyhydroxyethyl methacrylate is coated on the surface of activated carbon so that water-insoluble particles are not generated. That is, as one countermeasure against water-insoluble fine particles, the adsorbent is coated. However, since the adsorption material needs to be coated, the manufacturing process becomes complicated.

As mentioned above, from the viewpoint of the safety of water-insoluble particles, special processes are used to manufacture these adsorbers for direct blood perfusion. Since these special steps are required, the manufacturing cost of the product also increases. Therefore, there is a demand for an adsorber for direct blood perfusion that can be easily obtained using a general-purpose water-insoluble carrier without requiring special granulation equipment or coating equipment.

In addition, a patent for removing water-insoluble particles related to an adsorbent for medical use has been disclosed so far (see Patent Document 1), but it is characterized by having a plurality of circulation and particle removal lines, and there is no description of an adsorbent for removing water-insoluble particles. Particle size distribution, and there is no provision for the ratio of the mesh opening size for removing water-insoluble particles to the mesh opening size of the adsorber.

Patent Document 1: Japanese Unexamined Patent Publication No. 04-14594

Patent Document 2: Japanese Unexamined Patent Publication No. 01-275601

Patent Document 3: JP-A-62-191033

Non-Patent Document 1: Japan Medical Devices Industry Association, Guidelines for Specific Insurance Medical Materials, p175～, 2003

Non-Patent Document 2: Artificial Organs, Vol.23, No.2, p439～, 1994

Non-Patent Document 3: Chemical Engineering, Vol.50, No.10, p685～, 1986

Contents of the invention

The present invention provides and does not need special granulation equipment, coating equipment, uses the water-insoluble carrier (average particle diameter calculated on the basis of the number of particles to be 300～600 μm, particle size distribution) with relatively easy to obtain particle size distribution. The coefficient of variation is 10% to 20%), and the adsorber for direct blood perfusion of the adsorbent that removes the water-insoluble particles, and the method of obtaining the adsorbent for direct blood perfusion that removes the water-insoluble particles.

The inventors of the present invention have developed an adsorber for direct blood perfusion filled with an adsorbent that does not require special granulation equipment and coating equipment, uses a water-insoluble carrier having a relatively easily obtained particle size distribution, and removes water-insoluble particles. Obtained intensive research. As a result, the invention of an adsorber for direct blood perfusion filled with an adsorbent capable of removing water-insoluble fine particles at a therapeutically safe level without loss of the adsorbent when removing the water-insoluble fine particles has been completed.

That is, the present invention relates to an adsorber for direct blood perfusion filled with an adsorbent comprising water with an average particle diameter of 300 to 600 μm and a coefficient of variation of particle diameter distribution of 10% to 20%, calculated on the basis of the number of particles. Insoluble carrier particle group, the average +6SD of the measured value of the number of water-insoluble particles of 10 μm or more present in the adsorber per adsorber is 24,000 or less, and the measurement of the number of water-insoluble particles of 25 μm or more per adsorber The mean +6SD of the values is below 4000.

In addition, the present invention relates to the use of a sieve having a ratio of 1.3 to 1.5 between the mesh size of the mesh for removing water-insoluble fine particles and the mesh size of the adsorber. A method of obtaining an adsorbent filled in an adsorber for direct blood perfusion by removing water-insoluble fine particles from an adsorbent having a particle size distribution coefficient of variation of 10% to 20%.

According to the present invention, it is possible to obtain a special granulation device filled with an adsorbent material that does not need to be used to produce an adsorbent that is easy to remove water-insoluble particles and has a uniform particle size distribution, and it is relatively easy to obtain without performing a coating process for suppressing the generation of water-insoluble particles. A water-insoluble carrier with a particle size distribution, and an adsorbent material that removes water-insoluble particles to a safe level for treatment is an adsorber for direct blood perfusion. According to the present invention, an adsorber for direct blood perfusion is easily available, and the use of such an adsorber can also improve the safety of water-insoluble particles during treatment.

Description of drawings

Figure 1 is an overview of particle removal.

Figure 2 is the adsorber.

Figure 3 shows the relationship between flow and pressure loss.

Symbol Description

1 Adsorbent material

2 water insoluble particles

3 Screen to remove water-insoluble particles

4 filters

5 pumps

6 nozzles

7 Particle removal tank

8 liquid inlet

9 blood outflow port

10 columns

11 Adsorber

12 The screen of the adsorber

Detailed ways

Hereinafter, specific examples will be given to describe embodiments of the present invention.

The adsorbent in the present invention is not particularly limited as long as it has an affinity for the causative substance. As an example, an adsorbent in which a ligand having an affinity for a causative substance is immobilized on a water-insoluble carrier may be mentioned.

The water-insoluble carrier in the present invention is solid and water-insoluble at normal temperature and pressure, and has pores of appropriate size, that is, has a porous structure. As the water-insoluble carrier, spherical or granular carriers can be used. General-purpose carriers conventionally used in applications such as gel filtration agents, raw materials for ion exchangers, carriers for affinity chromatography, polymer carriers, carriers for body fluid purification, and cosmetic additives can be used.

In the direct blood perfusion method, when a spherical or granular water-insoluble carrier is used, in order to ensure a flow path through which blood cells and the like pass, the particle diameter must be larger than that of a liquid component such as blood plasma (plasma perfusion method). However, as the particle size becomes larger, the diffusion distance of the adsorbed pathogenic substance increases and the dynamic adsorption performance decreases. That is, the treatment time is prolonged. As the adsorbent in the present invention, it is preferable that the average particle diameter of the adsorbent capable of exhibiting good blood passage and adsorption efficiency in direct blood perfusion is 300-600 μm, more preferably 400-500 μm.

In the direct blood perfusion method, it is preferable that the particle size distribution of the water-insoluble carrier is more uniform from the viewpoint of hematocrit. However, granulation of carriers with uniform particle size distribution becomes difficult and requires special equipment. On the other hand, when the particle size distribution is widened, the adsorbent is tightly packed, and blood circulation becomes difficult because the flow path of blood becomes narrow. Furthermore, when the particle size distribution is wide, the amount of loss of the adsorbent accompanying the removal process increases when water-insoluble fine particles are removed.

As the adsorbent of the present invention, a general-purpose water-insoluble carrier having a particle size distribution that is relatively easy to obtain without requiring special granulation equipment, etc. can be used. The coefficient of variation of the particle size distribution (standard deviation calculated based on the number of particles in the sample/average particle size×100) is realistically preferably 10% to 20%. In addition, the average particle diameter of the adsorbent and the coefficient of variation of the particle diameter distribution are obtained by measuring the diameters of 100 or more particles one by one from magnified photographs of the adsorbent obtained with a stereo microscope, etc., based on the number of particles.

The strength of the water-insoluble carrier in the present invention is too soft and easy to be damaged, which is not preferable. If it is compacted when flowing through blood, sufficient blood flow cannot be obtained, and the treatment time cannot be extended and further treatment cannot be continued. In order to prevent the adsorption material The compaction of the adsorbent material is preferably an adsorbent material (hard) with sufficient mechanical strength. The so-called hard here, as shown in the reference example mentioned later, means that the adsorbent material is evenly filled into a cylindrical column and flowed through The relationship between the pressure loss and the flow rate of the aqueous liquid is a linear relationship at least up to 0.3kgf/cm ² . Also, a water-insoluble carrier that continuously generates water-insoluble fine particles during various steps such as transportation and filling is not preferable.

The material of the water-insoluble carrier in the present invention is not particularly limited, and representative examples include organic carriers made of polysaccharides such as cellulose, cellulose acetate, chitin, chitosan, dextrin, and agarose, polyphenylene Synthetic polymers such as ethylene, styrene-divinylbenzene copolymer, polyacrylamide, polyacrylic acid, polymethacrylic acid, polyacrylate, polymethacrylate, polyvinyl alcohol, etc. In addition, although it is not essential, it is also possible to have such a graft copolymerization in which a polymer material having a hydroxyl group such as polyhydroxyethyl methacrylate, a monomer having a polyethylene oxide chain, and other polymerizable monomers are copolymerized. coatings etc. Inorganic substances such as glass, alumina, and ceramics can also be used. Among them, synthetic polymers such as cellulose and polyvinyl alcohol are practically preferably used because active groups are easily introduced on the surface of the carrier.

Among them, a carrier made of cellulose is most preferably used. The carrier made of cellulose has relatively high mechanical strength and toughness, so it is less likely to be destroyed or produce water-insoluble particles. When filling the adsorber, it is difficult to compact even when blood flows through it at a high flow rate, so it can be used High velocity flows through the blood. In addition, it has the advantages of higher safety than synthetic polymer carriers, and is most suitable for use as the water-insoluble carrier in the present invention.

As an example of the etiological substance in the present invention, lipoproteins that are causative substances of arteriosclerosis, such as low specific gravity lipoproteins and ultra-low specific gravity lipoproteins present in body fluids, immunoglobulins (A.D.E.G.M), anti-DNA antibodies, Anti-acetylcholine receptor antibodies, anti-blood type antibodies, anti-platelet antibodies and other autoantibodies and antigen-antibody complexes, endotoxin, rheumatoid factor, macrophages, cancer tissue infiltrating T cells, etc.

The substance having an affinity for the causative substance in the present invention is not particularly limited as long as it adsorbs the target causative substance. The affinity between a substance having an affinity for a causative substance and a causative substance is divided into an affinity utilizing a biological interaction and a physicochemical interaction. Examples of the affinity utilizing biological interactions include affinity for immobilized antigens, affinity for immobilized antibodies, substances having affinity for causative substances utilizing complement fixation, Fc binding, and other biological interactions. As the affinity utilizing physical interaction, there are substances that have affinity for causative substances utilizing electrostatic interaction and hydrophobic interaction.

As a specific example of a substance having affinity for a causative substance utilizing physicochemical interactions, for example, a substance having an affinity for a causative substance having an anionic group can be used when low specific gravity lipoprotein is to be adsorbed. Examples of substances having an affinity for causative substances having negative groups include dextran sulfate, heparin, chondroitin sulfate, chondroitin polysulfate, heparan sulfate, xylan sulfate, and caronine sulfate. , cellulose sulfate, chitin sulfate, chitosan sulfate, pectin sulfate, inulin sulfate, alginic acid sulfate, glycogen sulfate, polylactose sulfate, carrageenan sulfate, starch sulfate, polydextrose sulfate, laminarin sulfate , galactan sulfate, fructan sulfate, mepesulfate and other sulfated polysaccharides, phosphotungstic acid, polysulfated anisole, polyvinyl alcohol sulfate, polyphosphoric acid, polyacrylic acid, etc. Among them, sulfated polysaccharides are particularly effective. In view of clinical practicality, preferred examples include heparin and dextran sulfate.

As a specific example of a substance that has affinity for a causative substance by utilizing hydrophobic interaction, when it is desired to adsorb fibrinogen, tryptophan derivatives, which are hydrophobic amino acids, can be used. The so-called tryptophan derivatives are Refers to tryptophan esters such as tryptophan, tryptophan ethyl ester, tryptophan methyl ester, tryptophan, tryptophanol (tryptophanol) and other compounds with a structure similar to tryptophan with an indole ring. In addition , these tryptophan derivatives can also be L-body, D-body, DL-body, or any of these mixtures. It can also be a mixture of two or more tryptophan derivatives. Among these tryptophan derivatives , in terms of safety, tryptophan is preferred, and among them, L-tryptophan is most preferably used practically because it is a natural amino acid, the data on its safety is abundant, cheap and easy to obtain.

Substances having affinity for various causative substances that exhibit interaction can be used depending on the target causative substance as exemplified above as substances having affinity for causative substances having a negative group and a hydrophobic group. In addition, it is also possible to immobilize a plurality of substances having an affinity for the aforementioned causative substances. As a specific example, when low specific gravity lipoprotein and fibrinogen are to be adsorbed at the same time, dextran sulfate and tryptophan derivatives may be simultaneously selected as substances having an affinity for the causative substance.

As a method of immobilizing a substance having an affinity for an etiological substance on a water-insoluble carrier in the present invention, there are known methods such as covalent bonding, ionic bonding, physical adsorption, embedding, precipitation insolubilization on the surface, etc. An appropriate method is selected for the material having affinity for the causative substance and the material of the above-mentioned water-insoluble carrier. In consideration of the eluting properties of substances having affinity for pathogenic substances during sterilization, immobilization and insolubilization by covalent bonds are preferred. In addition, a spacer may also be introduced between the water-insoluble carrier and the substance having an affinity for the causative substance as needed.

When a substance having an affinity for a causative substance is immobilized on a water-insoluble carrier by a covalent bond, the cyanogen halide method is known as an example of a method for increasing the reactivity of the water-insoluble carrier with a substance having an affinity for a causative substance , epichlorohydrin method, double epoxide method, bromoacetyl bromide method, etc. These methods are used to introduce functional groups such as amino group, carboxyl group, hydroxyl group, thiol group, acid anhydride group, succinimide group, chlorine group, aldehyde group, epoxy group and tresyl group on the water-insoluble carrier. Among them, an epoxy group derived by the epichlorohydrin method is particularly preferable in view of stability during heat sterilization.

The exclusion limit molecular weight of the globular protein of the water-insoluble carrier in the present invention must be larger than the molecular weight of the causative substance, and it is preferable to use a molecular weight of 2×10 ⁴ or more. The exclusion limit molecular weight means that, as described in monographs (Size Exclusion Chromatography, Moritada Yu, Kyoritsu Press), when samples with various molecular weights flow in Size Exclusion Chromatography, when they cannot penetrate into the pores (excluded) Intramolecular, the molecular weight of the substance with the smallest molecular weight. Specifically, for example, when the adsorbed pathogenic substance is low-density lipoprotein and blood fibrinogen, the adsorption capacity of fibrinogen and low-density lipoprotein is small when the exclusion limit molecular weight of globular protein is lower than 5×10 ⁵ , which is not practical. . On the other hand, the exclusion limit molecular weight of globular protein exceeds 1×10 ⁸ , the pore size (pore size) is too large, and the surface area available for adsorption decreases, resulting in a decrease in the adsorption capacity of fibrinogen and low-density lipoprotein. Therefore, the exclusion limit molecular weight of the globular protein of the water-insoluble carrier in the present invention is preferably 5×10 ⁵ to 1×10 ⁸ , more preferably 1×10 ⁶ to 1×10 ⁸ from the viewpoint of exhibiting adsorption performance, and even more preferably 2×10 ⁶ to 1×10 ⁸ .

In the adsorber of the present invention, the adsorbent of the present invention is filled in a column having a blood inlet and an outlet, and a mesh through which blood containing blood cell components passes but the adsorbent does not pass. In order to reduce the concentration of the causative substance in the blood, the capacity of the adsorber in the present invention must be 100 mL or more. From the standpoint of adsorption performance, there is no limit to the capacity of the adsorber, but the capacity of the adsorber is preferably 600 mL or less because there is a risk of lowering blood pressure if the amount of blood discharged outside the body is too large. In addition, even if it is incorporated into a circuit of other blood purification therapy such as hemodialysis, the amount of extracorporeal circulation of blood does not increase excessively, and from the viewpoint of preventing possible blood pressure drop that may occur as blood flows out of the body, it is preferable The capacity of the adsorber is below 400mL.

Since the adsorber of the present invention is used in the direct blood perfusion method which is easy to operate, the mesh of the adsorber must be a mesh so that the adsorbent does not flow out of the adsorber and must not catch blood cell components. When the mesh of the adsorber has a mesh smaller than that of the blood cell component is attached to the adsorber, the blood cell component is captured by the adsorber. On the other hand, when the mesh of the screen of the adsorber is larger than that of the adsorbent material, the adsorbent material in the adsorber flows out into the blood flow and enters the body. Therefore, the mesh size of the adsorber is preferably 100 to 200 μm.

The so-called water-insoluble particles in the present invention refer to water-insoluble particles that can flow out of the adsorber and enter the body without being captured by the screen of the adsorber during treatment. Therefore, the upper limit of the particle size defined as water-insoluble particles depends on The mesh size of the adsorber used. For example, when the mesh size of the adsorber is 150μm, the upper limit of the particle size of the water-insoluble particles is 150μm.

The presence of water-insoluble particulates of a size that can flow out of the adsorber in the adsorber is a big problem in terms of safety. According to the standard of insoluble particles in a certain injection (volume 100mL or more) described in the "Water-insoluble particle test method" of "No. 14 Corrected General Rules of Prescription Preparations 17 Injections", the following considerations may be related to the absorber for direct blood perfusion Standards for the safety of water-insoluble particulates.

There are two test methods described above, but in the second method "method using a microscope" which is more restrictive than the first method "method using a light-shielding automatic particle measuring device", the water insolubility of injections (volume 100mL or more) The standard of the number of particles is 12 or less water-insoluble particles of 100 μm or larger and 2 or less water-insoluble particles of 25 μm or larger in 1 mL. There are also cases where a large amount of 15-18 L is injected into the vein as a supplementary solution like hemofiltration, but as described in the monograph (Graphic Infusion Manual, written by Takeki Kitamura, published by Chinese and Foreign Medical Society), the total amount of infusion used as an infusion is usually 1 Day 2 ~ 2.5L or less. Here, 2L is taken as the total infusion volume. At this time, the maximum value allowed in the infusion solution is safe even if all the water-insoluble particles in the absorber enter the body during treatment. The indicator related to the safety of the number of water-insoluble particles is 10 μm or more. The number of water-insoluble fine particles is 24,000 or less per adsorber, and the number of water-insoluble fine particles of 25 μm or larger is 4,000 or less per adsorber. Therefore, as an indicator related to the safety of the number of water-insoluble particles, when considering the variation in the number of water-insoluble particles in each adsorber, the average +6SD of the measured values of the number of water-insoluble particles of 10 μm or more is 24,000 or less per adsorber, The average +6SD of the measured values of the number of water-insoluble fine particles of 25 μm or larger was 4000 or less per adsorber. Furthermore, the average and SD of the measured values of the number of water-insoluble particles in the present invention are the average value and standard deviation of the number of water-insoluble particles in each adsorber based on the number of particles measured by 6 adsorbers. . 6SD is 6 times the value of the standard deviation. The measurement of the number of water-insoluble particles in the present invention is carried out by the above-mentioned second method "method using a microscope", but it is also possible to use a Coulter counter diameter) were measured.

In the present invention, the number of water-insoluble fine particles is determined as follows. While passing water filtered through a 0.22 μm filter into the adsorber filled with the adsorbent, put the adsorbent in a container that does not generate water-insoluble particles (example: an empty container of commercially available saline solution) , transfer all the slurry to another prepared container, repeat this transfer operation 5 times between the containers to fully stir the adsorbent material, and then let the adsorbent material settle. Whether the adsorption material is settled or not, take the supernatant liquid, and use the above method to measure the particle concentration in the supernatant liquid. The number of water-insoluble fine particles contained in the adsorber is calculated from the measured value of the fine particle concentration to obtain the water-insoluble fine particles for each adsorber.

The removal of water-insoluble fine particles can also be performed by batch operation such as decantation, but it takes time and is inefficient. That is, it is preferable to continuously remove the water-insoluble fine particles as described below. Regarding the removal of water-insoluble fine particles in the present invention, a schematic diagram of an example is shown in FIG. 1 . In Fig. 1, 1 is an adsorption material, 2 is water-insoluble particles, 3 is a screen for removing particles, 4 is a filter, 5 is a pump, 6 is a nozzle, and 7 is a particle removal tank. The adsorbent is dispersed in the washing liquid in a container provided with a screen through which the water-insoluble fine particles pass but the adsorbent does not pass, and a filter having a filter for capturing the water-insoluble fine particles is used from the part where the adsorbent material separated by the screen does not exist Circulation and water-insoluble particle removal line, while using filter to capture water-insoluble particles in the washing liquid, inject the washing liquid through the nozzle, so that the adsorption material in the particle removal tank can be dispersed for a long time, and the water-insoluble particles can be removed.

FIG. 1 is merely an example, and it is not particularly limited as long as it has a screen for removing water-insoluble fine particles. In addition, there may be a plurality of lines for removing water-insoluble fine particles.

If the mesh size of the mesh for removing the water-insoluble particles is too small, the water-insoluble particles cannot be removed, and if it is too large, the adsorption material will flow out from the screen and cause loss when removing the water-insoluble particles. The filter of water-insoluble particles is clogged, and it is difficult to process continuously. The mesh size of the mesh for removing water-insoluble particles is preferably 1.3 to 1.5.

Next, a schematic cross-sectional view of an example of the adsorber of the present invention is shown in FIG. 2 . In Fig. 2, 1 is an adsorption material, 8 is an inflow port of blood, 9 is an outflow port of blood, 10 is a column, 12 is a screen of the adsorber, and 11 is an adsorber. However, the adsorber of the present invention is not limited to such a specific example, as long as it is an adsorber filled with an adsorbent material of a pathogenic substance in a container having a liquid inlet and an outlet and a mechanism to prevent the adsorbent from flowing out of the container, then The shape is not particularly limited.

Example

Hereinafter, a specific description will be given based on an embodiment of the present invention.

(reference example)

Agarose material (Biogel A-5m manufactured by Bio-rad, particle size 50-100 mesh) was uniformly filled in glass cylindrical columns (inner diameter 9 mm, column length 150 mm) equipped with filters with a pore size of 15 μm at both ends. , Vinyl-based polymer material (TOYOPEARL HW-65 manufactured by Tosoh Corporation, particle size 50-100 μm) and cellulose material (Cellulofine GC-700m manufactured by Chisso Corporation, particle size 45-105 μm), using a peristaltic pump to pass water , Find the relationship between the flow rate and the pressure loss ΔP. The results are shown in Fig. 3 .

As shown in Fig. 3, TOYOPEARL HW-65 and Cellulofine GC-700m increased the flow rate approximately proportional to the increase in pressure, while Biogel A-5m caused compaction, and the flow rate did not increase even when the pressure was increased. In the present invention, as in the former, the material whose relationship between the pressure loss ΔP and the flow rate is linear up to 0.3kgf/cm ² is called hard.

(manufacturing example)

Add 24L of porous cellulose beads with an average particle size of about 446 μm, a coefficient of variation of particle size distribution of 14%, and an exclusion limit molecular weight of globular proteins of 5×10 ⁷ , 6.6 L of 4N NaOH aqueous solution and 7 L of epichlorohydrin, and add water to make the total The volume was 33 L, and the reaction was carried out by stirring at 40°C for 2 hours. After the reaction, the beads were fully washed with water to obtain epoxidized cellulose beads. The epoxy group content of the epoxidized cellulose beads was 16.4 μmol/ml (wetted volume).

Prepare a dextran sulfate aqueous solution that dissolves 3 kg of dextran sulfate (sulfur content about 18%, molecular weight about 4000) in 15 L of water, add 24 L of epoxidized cellulose beads in a water-wet state, and use NaOH aqueous solution to make it alkaline Then, react at 45°C for 6 hours. After the reaction, the beads were sufficiently washed with water and saline, then a solution obtained by dissolving 0.34 kg of L-tryptophan in 11 L of dilute NaOH aqueous solution was added, and reacted at 50° C. for 8 hours. Then, the beads are fully washed with water and saline to obtain an adsorption material of dextran sulfate and tryptophan-immobilized cellulose beads. The average particle size of this adsorbent was 446 μm, and the coefficient of variation of the particle size distribution was 14%. In addition, the coefficient of variation of the average particle size and particle size distribution is to fully stir the slurry of the water-insoluble carrier and the adsorption material, take an appropriate amount and place it on a glass dish, and use a stereo microscope to focus on the adsorption material with a diameter of about 150 μm or more. Take the enlarged photo, measure 100 adsorbents by the 3-point circle method, and calculate based on the number of particles.

(Example 1)

The adsorption material of the production example was filled in an acrylic column (volume 2.7 mL) with an inner diameter of 10 mm and a length of 34 mm, and a polyethylene terephthalate screen with a mesh size of 150 μm was installed at both ends to obtain an adsorber. Add 5 units of heparin to 1 mL of blood, and circulate 40 mL of anticoagulated healthy human blood at a flow rate of 6.5 mL/min for 2 hours. The number of blood cells in the mixed blood before and after 2 hours of circulation is shown in Table 1. In addition, the concentrations of LDL-cholesterol, fibrinogen, and HDL-cholesterol in the mixed blood before and after circulation are shown in Table 2, and LDL-cholesterol is reduced from 163mg/dL to 105mg/dL dL, fibrinogen decreased from 210mg/dL to 152mg/dL, while HDL-cholesterol only decreased from 49mg/dL to 46mg/dL.

In addition, the amount of tryptophan immobilized on the adsorbent of the production example was obtained from the nitrogen content of the adsorbent. That is, 1 mL of the adsorbent in the production example was sufficiently washed with water, dried under reduced pressure at 60° C. for 6 hours or more, and then quantified using a trace total nitrogen analyzer. As a result, the amount of tryptophan immobilized on the adsorbent of the production example was 7.8 μmol/mL.

In addition, the amount of dextran sulfate immobilized on the adsorbent of the production example was measured by utilizing the affinity between dextran sulfate and toluidine blue. That is, about 100 mL of an aqueous solution of toluidine blue (basic blue 17 (Tokyo Chemical)) adjusted to about 90 mg/L was added to 3 mL of the adsorbent in the production example, stirred for 10 minutes, and after standing still, the absorbance at 630 nm was used. Quantify the above clarified toluidine blue, and obtain it from the amount of reduction. As a result, the amount of dextran sulfate immobilized on the adsorbent of the production example was 0.23 μmol/mL.

(Example 2)

The adsorbent of the production example (serum concentration 30%, liquid volume 16 L) was dispersed in a tank provided with a screen with a mesh size of 212 μm, and the slurry-solvent liquid (water) Use the circulation pipeline to inject the slurry-solvent solution into the tank through the nozzle, and disperse the adsorption material in the tank for a long time, while performing a 10-minute slurry-solvent circulation at a flow rate of 32L/min. In addition, in the setting A sieve with a mesh size of 104 μm was installed on the slurry-solvent circulation line outside the tank of the sieve, and the amount of the adsorbent that flowed out of the sieve with a mesh size of 122 μm and was captured by the sieve with a mesh size of 104 μm was obtained during the above-mentioned 10-minute cycle. The amount is shown in Table 3, which is 8.0mL.

Assuming that a 0.22 μm filter for capturing water-insoluble particles is installed instead of a 104-μm screen in the circulation line, and the water-insoluble particles are removed under the above conditions, it is estimated that the loss of the adsorbent accompanying the removal of the water-insoluble particles ( Outflow) amount is about 8.0 mL, which is small. In addition, if the amount of outflow is the same, clogging of the filter by the outflowing water-insoluble particles can be avoided by providing a pre-filter before the filter for capturing the water-insoluble particles during continuous operation.

In addition, when the mesh size of the adsorber is the same as in Example 1, and the mesh size of the water-insoluble particle removal tank is the same as that of the present example, the size of the mesh for removing water-insoluble particles (212 μm) is different from that of the mesh of the adsorber. The ratio of the size (150 μm) is 1.41.

(comparative example 1)

The adsorbent of the production example (serum concentration 30%, liquid volume 16 L) was dispersed in a tank provided with a screen with a mesh size of 233 μm, and the slurry-solvent liquid (water) Use the circulation pipeline to inject the slurry-solvent solution into the tank through the nozzle, and disperse the adsorbent material in the tank for a long time, and carry out the circulation of the slurry-solvent solution at a flow rate of 32L/min for 10 minutes. In addition, a sieve with a mesh size of 104 μm was installed on the slurry-solvent circulation line outside the tank where the mesh was installed, and the amount of water flowing out of the sieve with a mesh size of 233 μm and captured by a sieve with a mesh size of 104 μm was obtained during the above-mentioned 10-minute cycle. The amount of adsorbent material. This amount is shown in Table 3 and is 20 mL.

Assuming that a 0.22 μm filter for capturing water-insoluble particles is installed instead of a 104-μm screen in the circulation line, and the water-insoluble particles are removed under the above conditions, it is estimated that the loss of the adsorbent accompanying the removal of the water-insoluble particles ( Outflow) volume is about 20mL. This amount is more than Example 2. On the other hand, in order to avoid clogging of the filter for removing water-insoluble particles during continuous operation, even if a pre-filter is provided, the size of the pre-filter is larger than that of Example 2.

In addition, when the mesh size of the adsorber is the same as that of Example 1, and the mesh size of the water-insoluble particle removal tank is the same as that of this example, the mesh size (233 μm) of the mesh for removing water-insoluble particles is the same as that of the mesh size of the adsorber. The ratio of the mesh size (150 μm) is 1.55.

(Example 3)

The adsorbent of the production example (slurry concentration 15%, liquid volume 16L) was dispersed in the washing liquid in the particle removal tank provided with a screen with a mesh size of 212 μm, and from the part where the adsorbent separated by the screen did not exist, use a Circulation and water-insoluble particle removal line of 0.22 μm filter that captures water-insoluble particles flowing out of a 212 μm sieve, while using the filter to capture water-insoluble particles in the washing liquid, the washing liquid is circulated at a flow rate of 32 L/min. The nozzle re-injects the washing liquid into the particle removal tank, so that the adsorbent material in the particle removal tank is dispersed for a long time, and the water-insoluble particles are removed for 10 minutes. In addition, the mesh size (212μm) of the water-insoluble particles is removed and the adsorption device The ratio of mesh size (150μm) is 1.41.

In a closed system, about 250 mL of the adsorbent from which the water-insoluble fine particles were removed was filled into 6 columns having a screen of the adsorber used in Example 1 and a container of 250 mL, and then the adsorbent with a diameter of 0.22 μm was passed into the adsorber. The water edge filtered by the filter drives out the adsorbent material, and the number of water-insoluble particles in the adsorber is measured. (Since the mesh size of the adsorber is 150 μm, the upper limit of the particle size of the water-insoluble particles is 150 μm) For the determination method of the water-insoluble particles, refer to the water-insoluble particle test method of the 14th revision of the General Rules of the Prescription Preparations 17 Injection , Since the cellulose-based absorbent material is translucent, it cannot be measured by this method. Therefore, the water-insoluble fine particles were measured using a Coulter counter that measures the diameter equivalent to a sphere of equal volume by detecting the electrical resistance when the particles pass through the pores. However, since the water-insoluble carrier of the adsorbent is porous, the actual particle size is the value obtained by multiplying the particle size measured by a Coulter counter with a correction factor (actual particle size=measured particle size×2). (the measurement of the water-insoluble particle number of the following embodiment and comparative example adopts this measuring method to implement) the measurement result of the water-insoluble particle number in the adsorber of 6 adsorbers is as shown in table 4, in each adsorber 10～ The average +6SD of the measured values of the water-insoluble fine particles of 150 μm was 8474 pieces, and the average +6SD of the measured values of the water-insoluble fine particles of 25 to 150 μm in each adsorber was 1648 pieces. As mentioned above, the number of water-insoluble fine particles reached a safe level.

(Example 4)

The adsorbent of the production example (slurry concentration 30%, liquid volume 16 L) was dispersed in the washing liquid in the particle removal tank provided with a mesh of 212 μm, and from the part where the adsorbent separated by the mesh did not exist, a Circulation of 0.22 μm filter that captures water-insoluble particles flowing out of a 212 μm sieve · Water-insoluble particle removal line, while using the filter to capture water-insoluble particles in the washing liquid, the washing liquid is circulated at a flow rate of 32 L/min and passed through the nozzle The washing liquid is re-injected into the microparticle removal tank, and the adsorbent in the microparticle removal tank is dispersed for a long time, and the water-insoluble microparticles are removed for 10 minutes. In addition, the ratio of the mesh size (212 μm) for removing water-insoluble fine particles to the mesh size (150 μm) of the adsorber was 1.41.

In a closed system, about 250 mL of the adsorbent from which the water-insoluble fine particles were removed was filled into six 250-mL columns having the screen mesh of the adsorber used in Example 1, and a 0.22 μm The water filtered by the filter drives out the adsorbent material, and the number of water-insoluble particles in the adsorber is measured. (Because the mesh size of the adsorber is 150 μm, the upper limit of the particle size of the water-insoluble particles is 150 μm) The measurement results of the number of water-insoluble particles in the adsorbers of the six adsorbers are shown in Table 4. Each adsorber The average +6SD of the measured values of water-insoluble fine particles of 10 to 150 μm in the medium was 8501 pieces, and the average +6SD of the measured values of water-insoluble fine particles of 25 to 150 μm in each adsorber was 1652 pieces. As mentioned above, the number of water-insoluble fine particles reached a safe level.

(Example 5)

The adsorbent of the production example (slurry concentration 50%, liquid volume 6 L) was dispersed in the washing liquid in the particle removal tank provided with a screen with a mesh size of 212 μm, and from the part where the adsorbent separated by the screen did not exist, use a Circulation of 0.22 μm filter that captures water-insoluble particles flowing out of a 212 μm sieve · Water-insoluble particle removal line, while using the filter to capture water-insoluble particles in the washing liquid, the washing liquid is circulated at a flow rate of 32 L/min and passed through the nozzle Re-inject the cleaning solution into the particle removal tank to disperse the adsorbent material in the particle removal tank for a long time, and remove the water-insoluble particles for 10 minutes. In addition, the mesh size (212 μm) of the filter for removing water-insoluble particles is the same as the sieve of the adsorber. The ratio of mesh size (150μm) is 1.41.

In a closed system, about 250 mL of the adsorbent from which the water-insoluble fine particles were removed was filled into six 250-mL columns having the screen mesh of the adsorber used in Example 1, and a 0.22 μm The water filtered by the filter drives out the adsorbent material, and the number of water-insoluble particles in the adsorber is measured. (Because the mesh size of the adsorber is 150 μm, the upper limit of the particle size of the water-insoluble particles is 150 μm) The measurement results of the number of water-insoluble particles in the adsorbers of the six adsorbers are shown in Table 4. Each adsorber The average +6SD of the measured values of water-insoluble fine particles of 10 to 150 μm in the medium was 4412 pieces, and the average +6SD of the measured values of water-insoluble fine particles of 25 to 150 μm in each adsorber was 831 pieces. As mentioned above, the number of water-insoluble fine particles reached a safe level.

(comparative example 2)

The adsorbent of the production example (slurry concentration 30%, liquid volume 16 L) was dispersed in the washing liquid in the particle removal tank provided with a mesh of 180 μm, and the adsorbent separated by the mesh was used from the part where the adsorbent did not exist. Circulation of 0.22 μm filter that captures water-insoluble particles flowing out of a 180 μm sieve · Water-insoluble particle removal line, while using the filter to capture water-insoluble particles in the washing liquid, the washing liquid is circulated at a flow rate of 32 L/min and passed through the nozzle The washing solution is re-injected into the particle removal tank, and the adsorbent in the particle removal tank is dispersed for a long time, and the water-insoluble particles are removed for 10 minutes. In addition, the ratio of the mesh size (180 μm) for removing water-insoluble fine particles to the mesh size (150 μm) of the adsorber was 1.20.

In a closed system, about 250 mL of the adsorbent from which the water-insoluble fine particles were removed was filled into six 250-mL columns having the screen mesh of the adsorber used in Example 1, and a 0.22 μm The water filtered by the filter drives out the adsorbent material, and the number of water-insoluble particles in the adsorber is measured. (Since the mesh size of the adsorber is 150 μm, the upper limit of the particle size of the water-insoluble fine particles is 150 μm). Table 4 shows the measurement results of the number of water-insoluble fine particles in the adsorbers of the six adsorbers. The average +6SD of the measured values of water-insoluble fine particles of 10-150 μm in each adsorber was 23,909, and the average +6SD of measured values of water-insoluble fine particles of 25-150 μm in each adsorber was 13,142. As mentioned above, the number of water-insoluble fine particles did not reach a safe level.

Claims (5)

1. direct hemoperfusion adsorber, it is the direct hemoperfusion adsorber that is filled with adsorbing material, this adsorbing material is to obtain by following method: the ratio that uses the screen cloth eye size remove water insoluble microparticle and the screen cloth eye size of adsorber is 1.3～1.5 screen cloth, is that mean diameter that benchmark is calculated is that the coefficient of alteration of 300 μ m～600 μ m and particle size distribution is to remove water insoluble microparticle the adsorbing material of 10%～20% population from comprising with the particle number;

It is that mean diameter that benchmark is calculated is that the coefficient of alteration of 300 μ m～600 μ m, particle size distribution is the population of 10%～20% water insoluble carrier that this adsorbing material comprises with the particle number, the standard deviation of meansigma methods+6 of the measured value that is present in the above water insoluble microparticle number of 10 μ m in the adsorber of each adsorber times is below 24000, and the standard deviation of meansigma methods+6 of the measured value of the water insoluble microparticle number that 25 μ m of each adsorber are above times is below 4000.

2. the described direct hemoperfusion adsorber of claim 1, wherein, the exclusion limit molecular weight of the globular protein of adsorbing material is 2 * 10 ⁴～1 * 10 ⁸

3. claim 1 or 2 described direct hemoperfusion adsorbers, wherein, adsorbing material is the cellulose family carrier.

4. the described direct hemoperfusion adsorber of claim 1, wherein, adsorbing material is the cellulose family carrier, the exclusion limit molecular weight of the globular protein of adsorbing material is 5 * 10 ⁵～1 * 10 ⁸, and filled the adsorbing material of low density lipoprotein, LDL and fibrinogen.

5. obtain the method for direct hemoperfusion that be filled into the adsorbing material in the adsorber, wherein, the screen cloth eye size that water insoluble microparticle is removed in use and the ratio of the screen cloth eye size of adsorber are 1.3～1.5 screen cloth, are that mean diameter that benchmark is calculated is that the coefficient of alteration of 300 μ m～600 μ m and particle size distribution is to remove water insoluble microparticle the adsorbing material of 10%～20% population from comprising with the particle number.

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